Intervention (R2 = 0.4123, p = 3.13.1029 and R2 = 0.4185, p = 2.56.1029; Figs. 1A and B). This correlation came out to be even better, when the CD14 and THBD mRNA expression levels were further normalized by their respective VDR mRNA levels (R2 = 0.6709, p = 2.49.10211 and R2 = 0.7428, p = 1.88.10215; Figs. S3A and B in File S1). For all 71 individuals the change of mRNA expression levels of CD14 and THBD positively correlated to each other (R2 = 0.1479, p = 0.00099; Fig. 2A). Interestingly, an even far more significant correlation between the Linolenic acid methyl ester changes of CD14 and THBD mRNA expression levels was observed in adipose tissue samples obtained from 47 15481974 of the study participants (R2 = 0.5978, p = 3.10210; Fig. 2B). Taken together, the expected negative correlation between serum concentrations of PTH and 25(OH)D3 confirmed an overall normal physiological vitamin D response of the 71 study participants. The primary VDR target genes CD14 and THBD showed to be suitable transcriptomic biomarkers of vitamin D signaling both in PBMCs and adipocytes.CD14 and THBD mRNA expression changes efficiently classify study participantsBased on the proven overall functionality of vitamin D signaling in the tissue samples of the study participants we assumed that an buy Fruquintinib increase in serum 25(OH)D3 concentrations would result in a respective increase in CD14 and THBD mRNA expression. However, neither with all 71 PBMCs donors nor with the subgroup of all 47 adipose tissue donors we could observe any significant correlation between the changes in 25(OH)D3 serum concentrations and the mRNA expression changes of CD14 or THBD in the respective tissue samples (data not shown). Based on the assumption of a linear positive correlation between the changes of CD14 or THBD gene expression both in PBMCs and adipose tissue and changes of 25(OH)D3 serum concentrations, we performed four separate rankings of the study participants, which were then combined to one ranking (Table S2 in File S2).When we took only the top half of the ranking (35 persons), the plot of the changes in CD14 (Fig. 3A) or THBD (Fig. 3B) mRNA expression levels in PBMCs against the respective changes in 25(OH)D3 concentrations provided statistically significant positive correlations (R2 = 0.168, p = 0.01562 and R2 = 0.243, p = 0.00324). We could confirm this observation with the adipose tissue samples, when restricting to the top half of the ranking in this tissue (23 persons). However, in adipose tissue the positive correlation between the changes in CD14 (R2 = 0.1802, p = 0.02163; Fig. 3C) or THBD (R2 = 0.1723, p = 0.08199; Fig. 3D) mRNA expression levels and their respective 25(OH)D3 serum concentrations is less significant than in PBMCs. As a negative control for VDR target gene specificity, in neither of the two tissues the changes in VDR mRNA expression correlated with 25(OH)D3 serum concentration changes (data not shown). The categorization of the study participants by response to a change in their 25(OH)D3 serum concentrations allows to observe correlations that are not visible when all participants are included in the analyses. For example, on the basis of all 71 participants there 23977191 is no correlation between the change in serum concentrations of 25(OH)D3 and the inflammatory marker IL6 [41] (data not shown). However, when we analyzed the segregated set of the 35 most vitamin D responsive samples, we found a significant negative correlation between changes in 25(OH)D3 and IL6 (R2 = 0.1352, p = 0.03; Fig. 4.Intervention (R2 = 0.4123, p = 3.13.1029 and R2 = 0.4185, p = 2.56.1029; Figs. 1A and B). This correlation came out to be even better, when the CD14 and THBD mRNA expression levels were further normalized by their respective VDR mRNA levels (R2 = 0.6709, p = 2.49.10211 and R2 = 0.7428, p = 1.88.10215; Figs. S3A and B in File S1). For all 71 individuals the change of mRNA expression levels of CD14 and THBD positively correlated to each other (R2 = 0.1479, p = 0.00099; Fig. 2A). Interestingly, an even far more significant correlation between the changes of CD14 and THBD mRNA expression levels was observed in adipose tissue samples obtained from 47 15481974 of the study participants (R2 = 0.5978, p = 3.10210; Fig. 2B). Taken together, the expected negative correlation between serum concentrations of PTH and 25(OH)D3 confirmed an overall normal physiological vitamin D response of the 71 study participants. The primary VDR target genes CD14 and THBD showed to be suitable transcriptomic biomarkers of vitamin D signaling both in PBMCs and adipocytes.CD14 and THBD mRNA expression changes efficiently classify study participantsBased on the proven overall functionality of vitamin D signaling in the tissue samples of the study participants we assumed that an increase in serum 25(OH)D3 concentrations would result in a respective increase in CD14 and THBD mRNA expression. However, neither with all 71 PBMCs donors nor with the subgroup of all 47 adipose tissue donors we could observe any significant correlation between the changes in 25(OH)D3 serum concentrations and the mRNA expression changes of CD14 or THBD in the respective tissue samples (data not shown). Based on the assumption of a linear positive correlation between the changes of CD14 or THBD gene expression both in PBMCs and adipose tissue and changes of 25(OH)D3 serum concentrations, we performed four separate rankings of the study participants, which were then combined to one ranking (Table S2 in File S2).When we took only the top half of the ranking (35 persons), the plot of the changes in CD14 (Fig. 3A) or THBD (Fig. 3B) mRNA expression levels in PBMCs against the respective changes in 25(OH)D3 concentrations provided statistically significant positive correlations (R2 = 0.168, p = 0.01562 and R2 = 0.243, p = 0.00324). We could confirm this observation with the adipose tissue samples, when restricting to the top half of the ranking in this tissue (23 persons). However, in adipose tissue the positive correlation between the changes in CD14 (R2 = 0.1802, p = 0.02163; Fig. 3C) or THBD (R2 = 0.1723, p = 0.08199; Fig. 3D) mRNA expression levels and their respective 25(OH)D3 serum concentrations is less significant than in PBMCs. As a negative control for VDR target gene specificity, in neither of the two tissues the changes in VDR mRNA expression correlated with 25(OH)D3 serum concentration changes (data not shown). The categorization of the study participants by response to a change in their 25(OH)D3 serum concentrations allows to observe correlations that are not visible when all participants are included in the analyses. For example, on the basis of all 71 participants there 23977191 is no correlation between the change in serum concentrations of 25(OH)D3 and the inflammatory marker IL6 [41] (data not shown). However, when we analyzed the segregated set of the 35 most vitamin D responsive samples, we found a significant negative correlation between changes in 25(OH)D3 and IL6 (R2 = 0.1352, p = 0.03; Fig. 4.
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